Disease Information: Entamoeba Histolytica is the pathogenic amoeba which causes amebiasis, a parasitic infection which gives rise to (extra)intestinal diseases. Amebiasis is spread by fecal contamination (water, soil, improper sanitation) as well as through fecal-oral transmission during rectal inoculation or sexual exchange. An estimated 50 million cases occur every year, mainly in developing countries. Amebiasis can be asymptomatic, and it is predicted that as high as 25% of individuals in endemic places carry the amoeba antibodies. Though it affects individuals of all ages, mainly adult men are impacted by invasive amebiasis (it is suggested that this may be the result of differences in endocrinological processes between the sexes). A link between HIV/AIDS and amebiasis has also been made but the epidemiological impact of one on the other is disputed at this point in time.
Essentiality of this protein: This protein reduces thioredoxin, which allows for correct folding of target proteins, and also reduces hydrogen peroxide (which is toxic to the organism).
Complex of proteins?: Multimer ("2-mer")
Druggable Target (list number or cite evidence from a paper/database showing druggable in another organism): See this list
Image of protein (PyMol with features delineated and shown separately): *Amino Acid Sequence (paste as text only - not as screenshot or as 'code')
>4CCQ:A|PDBID|CHAIN|SEQUENCE
*GC% Content for gene: *CDS Gene Sequence (codon optimized) - copy fromoutputofPrimer DesignProtocol (paste as text only): *GC% Content for gene (codon optimized):
Do Not Need this info for Spring (but still copy these lines to your Target page for now) Primer design results for pNIC-Bsa4 cloning (list seqeunces of all of your ~40 nt long primers): (link to DNA Works output text file - that should be saved in your Google Docs folder after you did the primer design protocol) -- Ask a mentor, Dr. B, or a fellow researcher -how to link a GDocs file if you are not sure how to.
Primer design results for 'tail' primers (this is just 2 sequences): **
*NCBI Gene # or RefSeq#: 3410063
*Protein ID (NP or XP #) or Wolbachia#: EHI_155440
*Organism (including strain): Entamoeba Histolytica
Etiologic Risk Group (see link below): 2
Disease Information:
Entamoeba Histolytica is the pathogenic amoeba which causes amebiasis, a parasitic infection which gives rise to (extra)intestinal diseases. Amebiasis is spread by fecal contamination (water, soil, improper sanitation) as well as through fecal-oral transmission during rectal inoculation or sexual exchange. An estimated 50 million cases occur every year, mainly in developing countries. Amebiasis can be asymptomatic, and it is predicted that as high as 25% of individuals in endemic places carry the amoeba antibodies. Though it affects individuals of all ages, mainly adult men are impacted by invasive amebiasis (it is suggested that this may be the result of differences in endocrinological processes between the sexes). A link between HIV/AIDS and amebiasis has also been made but the epidemiological impact of one on the other is disputed at this point in time.
Gene database link here
Essentiality of this protein:
This protein reduces thioredoxin, which allows for correct folding of target proteins, and also reduces hydrogen peroxide (which is toxic to the organism).
Complex of proteins?: Multimer ("2-mer")
Druggable Target (list number or cite evidence from a paper/database showing druggable in another organism):
See this list
*EC#: 1.8.1.9
Link to BRENDA EC# page: Brenda Page
Enzyme Assay information (spectrophotometric, coupled assay ?, reagents):
Assay Kit (an E.Coli version is also available)
Substrate
NADP: N8035 SIGMA (5 mg per vial) = $174.00
6/6/2016 Edit: DTNB
Structure (PDB or Homology model)
PDB#: 4A5L
Current Inhibitors:
Azelaic Acid in Staphylococcus aureus (strain Mu50)
Expression Information (has it been expressed in bacterial cells): Yes, through E.Coli, engineered with a 6-His Tag
Purification Method: Ni-NTA affinity column chromatography
Image of protein (PyMol with features delineated and shown separately):
*Amino Acid Sequence (paste as text only - not as screenshot or as 'code')
>4CCQ:A|PDBID|CHAIN|SEQUENCE
MSNIHDVVIIGSGPAAHTAAIYLGRSSLKPVMYEGFMAGGVAAGGQLTTTTIIENFPGFPNGIDGNELMMNMRTQSEKYG
TTIITETIDHVDFSTQPFKLFTEEGKEVLTKSVIIATGATAKRMHVPGEDKYWQNGVSACAICDGAVPIFRNKVLMVVGG
GDAAMEEALHLTKYGSKVIILHRRDAFRASKTMQERVLNHPKIEVIWNSELVELEGDGDLLNGAKIHNLVSGEYKVVPVA
GLFYAIGHSPNSKFLGGQVKTADDGYILTEGPKTSVDGVFACGDVCDRVYRQAIVAAGSGCMAALSCEKWLQTH
>4CCQ:B|PDBID|CHAIN|SEQUENCE
MSNIHDVVIIGSGPAAHTAAIYLGRSSLKPVMYEGFMAGGVAAGGQLTTTTIIENFPGFPNGIDGNELMMNMRTQSEKYG
TTIITETIDHVDFSTQPFKLFTEEGKEVLTKSVIIATGATAKRMHVPGEDKYWQNGVSACAICDGAVPIFRNKVLMVVGG
GDAAMEEALHLTKYGSKVIILHRRDAFRASKTMQERVLNHPKIEVIWNSELVELEGDGDLLNGAKIHNLVSGEYKVVPVA
GLFYAIGHSPNSKFLGGQVKTADDGYILTEGPKTSVDGVFACGDVCDRVYRQAIVAAGSGCMAALSCEKWLQTH
Length of your protein in Amino Acids : 649
Molecular Weight: 69827.7 kD
Molar Extinction coefficient of your protein at 280 nm wavelength: 59820 M^-1 cm^-1
TMpred graph Image
*CDS Gene Sequence (paste as text only):
ATGAGTAATATTCATGATGTTGTGATTATCGGCTCAGGACCTGCTGCACATACTGCGGCTATTTATTTAG
GAAGATCTTCATTGAAACCAGTTATGTATGAAGGGTTTATGGCAGGAGGAGTAGCTGCAGGAGGACAACT
AACTACCACTACTATCATTGAGAATTTCCCAGGATTTCCAAACGGTATTGATGGAAATGAATTAATGATG
AATATGAGAACACAATCAGAGAAGTATGGAACTACTATCATTACAGAAACTATTGACCATGTTGACTTTT
CAACACAACCATTTAAATTGTTTACTGAAGAAGGAAAAGAGGTACTTACAAAATCAGTTATTATTGCAAC
AGGTGCTACAGCTAAAAGAATGCATGTTCCAGGAGAAGACAAATATTGGCAAAATGGAGTTAGTGCATGT
GCCATTTGTGATGGGGCTGTTCCTATTTTCAGAAATAAAGTATTAATGGTAGTTGGTGGAGGAGATGCAG
CTATGGAAGAAGCACTTCATTTAACAAAATATGGAAGTAAAGTTATTATTCTTCATCGAAGAGATGCTTT
TAGGGCAAGTAAAACTATGCAAGAACGAGTATTAAATCATCCAAAGATTGAAGTAATATGGAATTCTGAA
TTAGTAGAATTAGAAGGAGATGGTGATTTATTAAATGGGGCAAAAATTCATAATCTAGTTTCAGGAGAAT
ACAAAGTTGTTCCAGTAGCTGGATTGTTTTATGCTATTGGACATAGTCCAAATAGTAAATTCTTAGGTGG
ACAAGTTAAAACTGCTGATGATGGATATATTCTTACAGAAGGACCAAAAACTAGTGTTGATGGTGTATTT
GCATGTGGGGATGTATGTGATAGAGTATATAGACAAGCTATTGTAGCTGCAGGAAGTGGTTGTATGGCAG
CCTTAAGCTGTGAAAAATGGCTTCAAACTCATTAATTCAGTTTCTTTTTTAACA
*GC% Content for gene:
*CDS Gene Sequence (codon optimized) - copy from output of Primer Design Protocol (paste as text only):
*GC% Content for gene (codon optimized):
Do Not Need this info for Spring (but still copy these lines to your Target page for now)
Primer design results for pNIC-Bsa4 cloning (list seqeunces of all of your ~40 nt long primers):
(link to DNA Works output text file - that should be saved in your Google Docs folder after you did the primer design protocol)
-- Ask a mentor, Dr. B, or a fellow researcher -how to link a GDocs file if you are not sure how to.
Primer design results for 'tail' primers (this is just 2 sequences):
**